Choline Supplementation Normalizes Fetal Adiposity and Reduces Lipogenic Gene Expression in a Mouse Model of Maternal Obesity

Chauntelle Jack-Roberts, Yaelle Joselit, Khatia Nanobashvili, Rachel Bretter, Olga V Malysheva, Marie A Caudill, Anjana Saxena, Kathleen Axen, Ahmed Gomaa, Xinyin Jiang, Chauntelle Jack-Roberts, Yaelle Joselit, Khatia Nanobashvili, Rachel Bretter, Olga V Malysheva, Marie A Caudill, Anjana Saxena, Kathleen Axen, Ahmed Gomaa, Xinyin Jiang

Abstract

Maternal obesity increases fetal adiposity which may adversely affect metabolic health of the offspring. Choline regulates lipid metabolism and thus may influence adiposity. This study investigates the effect of maternal choline supplementation on fetal adiposity in a mouse model of maternal obesity. C57BL/6J mice were fed either a high-fat (HF) diet or a control (NF) diet and received either 25 mM choline supplemented (CS) or control untreated (CO) drinking water for 6 weeks before timed-mating and throughout gestation. At embryonic day 17.5, HF feeding led to higher (p < 0.05) percent total body fat in fetuses from the HFCO group, while the choline supplemented HFCS group did not show significant difference versus the NFCO group. Similarly, HF feeding led to higher (p < 0.05) hepatic triglyceride accumulation in the HFCO but not the HFCS fetuses. mRNA levels of lipogenic genes such as Acc1, Fads1, and Elovl5, as well as the transcription factor Srebp1c that favors lipogenesis were downregulated (p < 0.05) by maternal choline supplementation in the HFCS group, which may serve as a mechanism to reduce fat accumulation in the fetal liver during maternal HF feeding. In summary, maternal choline supplementation improves indices of fetal adiposity in obese dams at late gestation.

Keywords: choline; fetal adiposity; gestation; lipogenesis; obesity.

Conflict of interest statement

The authors declare no conflicts of interest. The founding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in the decision to publish the results.

Figures

Figure 1
Figure 1
Design of the study. Female C57BL/6J mice were divided into four groups and fed the normal fat (NF) normal choline (CO) diet, NF choline supplemented (CS) diet, HFCO diet, or HFCS diet for 6 weeks before timed-mating and throughout gestation. Male mice followed the NFCO diet until timed-mating. The intraperitoneal glucose tolerance test (GTT) was conducted at embryonic day (E) 15.5 and dissection was conducted at E17.5. n is the number of dams in each group from which data and samples were collected.
Figure 2
Figure 2
Weight gain and intraperitoneal glucose tolerance (IGT) of dams fed experimental diets. (a) Weight gain was measured both before timed-mating and during gestation (E0.5–E17.5); (b) Visceral fat was measured at E17.5; (c) IGT tests were conducted at E15.5 with 2 g/kg D-glucose injected; (d) The area under the curve (AUC) of the IGT tests. NFCO (solid bars): n = 10; NFCS (shaded bars): n = 6; HFCO (open bars): n = 8; HFCS (hatched bars): n = 7. Data were analyzed using the general linear model. Posteriori analysis of variance (ANOVA) followed by posthoc Fisher‘s least significant difference (LSD) analysis between groups was conducted with pd×s ≤ 0.2. Values are mean ± standard error of mean (SEM); different letters indicate p < 0.05 in the posthoc analysis. ns: pd, ps, and pd×s, not significant. CO: control; CS: choline supplemented; D: diet; HF: high-fat diet; NF: normal-fat diet; S: supplementation.
Figure 3
Figure 3
Fetal weight and adiposity at E17.5. (a) Fetal weight; (b) percent total body fat of fetuses; (c) fetal liver triglyceride concentrations. Experimental diets were fed to dams from 6 weeks before timed-mating to gestational day 17.5. Data were analyzed using the general linear model. Posteriori analysis of variance (ANOVA) followed by posthoc Fisher‘s least significant difference (LSD) analysis between groups was conducted with pd×s ≤ 0.2. NFCO (solid bars): n = 9; NFCS (shaded bars): n = 6; HFCO (open bars): n = 7; HFCS (hatched bars): n = 6. n is the number of dams. Fetal weight was analyzed with pooled data of all fetuses in each dam (5–10/dam); body fat and liver triglyceride concentrations were analyzed with pooled data of two randomly selected fetuses in each dam. Values are mean ± standard error of mean (SEM); different letters indicate p < 0.05 in the posthoc analysis. ns: pd, pS, and pd×s, not significant. CO: control; CS: choline supplemented; D: diet; HF: high-fat diet; NF: normal-fat diet; S: supplementation.
Figure 4
Figure 4
Fetal liver mRNA abundance at E17.5. Experimental diets were fed to dams from 6 weeks before timed-mating to gestational day 17.5. (a) Genes involved in lipid metabolism; (b) transcription factors; (c) genes involved in choline metabolism. mRNA levels were measured by real-time PCR. Data were analyzed using the general linear model. Data were analyzed using the general linear model. Posteriori analysis of variance (ANOVA) followed by posthoc Fisher's least significant difference (LSD) analysis between groups was conducted with pd×s ≤ 0.2. NFCO (solid bars): n = 9; NFCS (shaded bars): n = 6; HFCO (open bars): n = 7; HFCS (hatched bars): n = 6. n is the number of dams. Data from two fetuses in each dam were pooled and included in the analysis. Values are mean ± standard error of mean (SEM); different letters indicate p < 0.05 in the posthoc analysis; ns: pd, pS, and pd×s, not significant. Acc: acetyl-CoA carboxylase; Acox1: peroxisomal acyl-coenzyme A oxidase 1; Bhmt: betaine-homocysteine S-methyltransferase; Chdh: choline dehydrogenase; Chrebp1: Carbohydrate-responsive element-binding protein; Dgat1: diacylglycerol O-acyltransferase 1; Elovl5: fatty acid elongase 5; Fasn: fatty acid synthase; Fatp1: fatty acid transport protein 1; Mttp: microsomal triglyceride transfer protein; Pcyt1a: choline-phosphate cytidylyltransferase A; Pemt: phosphatidylethanolamine N-methyltransferase; Ppara: Peroxisome proliferator-activated receptor alpha; Scd1: stearoyl-CoA desaturase-1; Srebp1: Sterol regulatory element-binding protein 1; CO: control; CS: choline supplemented; D: diet; HF: high-fat diet; NF: normal-fat diet; S: supplementation.

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